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Apoptosis and Necrosis Quantification Kit

A convenient assay for quantifying apoptotic (green) and necrotic (red) cells within the same cell population by flow cytometry or fluorescence microscopy

Product Attributes

Apoptosis/viability marker

Phosphatidylserine/Annexin V, Dead cell stain, Apoptosis/necrosis assay

For live or fixed cells

For live/intact cells

Detection method/readout

Fluorescence microscopy, Flow cytometry

Assay type/options

Endpoint assay

Colors

Green/Red

Size
Catalog #
price
Qty
50 assays
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Product Description

The Apoptosis and Necrosis Quantitation Kit provides a convenient assay for quantifying apoptotic (green) and necrotic (red) cells within the same cell population by flow cytometry or fluorescence microscopy.

Apoptosis and necrosis are two major processes by which cells die. Apoptosis is an active, genetically regulated disassembly of the cell from within. During apoptosis, phosphatidylserine (PS) is translocated from the inner to the outer leaflet of the plasma membrane for phagocytic cell recognition. The human anticoagulant Annexin V is a 35 kDa, Ca2 -dependent phospholipid-binding protein with high affinity for PS. Annexin V labeled with fluorescein (FITC) stains apoptotic cells green by binding to PS exposed on the cell surface.

Necrosis normally results from severe cellular insult. Both internal organelle and plasma membrane integrity are lost, resulting in the release of cellular contents into the surrounding environment. Ethidium homodimer III (EthD-III) is a highly positively charged nucleic acid probe, which is impermeant to live cells, but stains cells with compromised membrane integrity with red fluorescence. EthD-III is a superior alternative to propidium iodide (PI) or ethidium homodimer I because of its significantly higher affinity for DNA and higher fluorescence quantum yield. The Apoptosis & Necrosis Quantitation Kit provides a convenient assay for quantifying apoptotic (green) and necrotic (red) cells within the same cell population by flow cytometry or fluorescence microscopy.

Features:

  • Simple: Simultaneous detection of both apoptotic and necrotic cells in a single assay
  • Fast: A single 15-30 min staining step
  • Sensitive: EthD-III is a superior alternative to both PI and EthD-I for staining necrotic cells.

Kit Components:

  • FITC-Annexin V
  • Ethidium homodimer III
  • 5X binding buffer

Spectral properties (Ex/Em):

  • Fluorescein (FITC): 498/517 nm
  • Ethidium homodimer III (EthD-III) (with DNA): 522/593 nm (EthD-III also has a large absorption peak at 279 nm)

Also see the Apoptosis and Necrosis Quantitation Kit Plus (catalog no. 30065) and Apoptotic, Necrotic, and Healthy Cells Quantitation Kit Plus (catalog no. 30066), which feature Annexin V conjugated to Biotium’s superior green fluorescent dye CF®488A, which is brighter and more photostable than FITC.

References

1. BMC Immunology (2010) doi:10.1186/1471-2172-10-29.

2. Biosci Rep 7, 289 (1987).

3. FEBS Lett 203, 99 (1986).

 

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